牙龈卟啉单胞菌通过干扰肠道菌群对糖尿病小鼠血糖的影响

doi:10.13701/j.cnki.kqyxyj.2023.11.013

口腔医学研究 ›› 2023, Vol. 39 ›› Issue (11): 1005-1011.DOI: 10.13701/j.cnki.kqyxyj.2023.11.013

牙龈卟啉单胞菌通过干扰肠道菌群对糖尿病小鼠血糖的影响

陈文文¹, 闫福华², 莫朝伦³, 黄明坤¹, 李萧纹¹, 张军梅^3*

1.贵州医科大学口腔医学院贵州贵阳 550004;
2.南京大学医学院附属口腔医院牙周病科江苏南京 210008;
3.贵州医科大学附属口腔医院正畸二科贵州贵阳 550001

收稿日期:2023-05-30 出版日期:2023-11-28 发布日期:2023-11-22
通讯作者: ^*张军梅,E-mail:zjm46688@126.com
作者简介:陈文文(1997~ ),女,贵州福泉人,硕士在读,主要从事牙周炎与糖尿病相关关系及肠道营养的基础和临床研究。
基金资助:
国家自然基金项目(编号:81970939)贵州省卫生健康委项目(编号:gzwkj2023-61)

Effect of Porphyromonas Gingivalis on Blood Glucose and Intestinal Flora in Diabetic Mice: A Mechanistic Exploration

CHEN Wenwen¹, YAN Fuhua², MO Chaolun³, HUANG Mingkun¹, LI Xiaowen¹, ZHANG Junmei^3*

1. School of Stomatology, Guizhou Medical University, Guiyang 550004, China;
2. Department of Periodontology, Nanjing Stomatological Hospital, Medical School of Nanjing University, Nanjing 210008, China;
3. Department of the Second Orthodontic, Affiliated Stomatological Hospital of Guizhou Medical University, Guiyang 550001, China

Received:2023-05-30 Online:2023-11-28 Published:2023-11-22

摘要/Abstract

摘要： 目的: 探究牙龈卟啉单胞菌(Porphyromonas gingivalis,P.g)对糖尿病(diabetes,DM)小鼠血糖的影响,并在肠道菌群中筛选出与血糖密切相关的菌种,初步探索此干预的相关机制。方法: 将32只野生型C57BL/6J小鼠按照随机对照重复盲法原则分为4组(每组8只):健康小鼠+灌饲PBS(WT组),健康小鼠+灌饲P.g组(WT P.g组),糖尿病+灌饲PBS组(STZ组),糖尿病+灌饲P.g组(STZP.g组)。1周1次观察小鼠血糖、体重及精神状态,采用16S rDNA检测小鼠结肠粪便内肠道菌群变化,酶联免疫吸附试验及免疫荧光技术检测结肠段胰高血糖素样肽(glucagon like peptide-1,GLP-1)的表达差异,q-PCR测定结肠组织中前激素转化酶1/3(proconvertase 1/3,PC1/3)和胰高血糖素原(proglucagon)mRNA的表达。结果: 与WT组相比,WT P.g组空腹血糖(fasting blood glucose,FBG)无差别;而与STZ组相比,灌胃第6周时STZ P.g组差异有统计学意义(P<0.05),且肠道菌群丰富度及多样性明显降低。结果显示STZ P.g组肠L细胞明显遭受破坏,q-PCR测定结果显示PC1/3、proglucagon mRNA异常表达,并免疫荧光提示GLP-1表达量明显减低且有统计学差异(P<0.05)。经16S rDNA结果显示,在属水平上,STZ P.g组气单胞菌属(Aeromonas)、罗斯氏菌属(Rothia)、副普雷沃氏菌属(Paraprevotella)、梭菌属(Clostridium)显著性增加,宿主关联乳杆菌属(Ligilactobacillus)、脱硫弧菌属(Desulfovibrio)、阿克曼氏菌属(Akkermansia)、拟杆菌属(Bacteroides)和毛罗菌_NK4A136_群属(Lachnospiraceae_NK4A136_group)显著性减少。FBG与Lachnospiraceae_NK4A136_group、肠杆菌属(Enterorhabdus)和念珠菌(Candidatus_Saccharimonas)丰富度呈明显负相关,而与Desulfovibrio、Akkermansia、Bacteroides呈明显正相关。结论: P.g可能通过肠道菌群干预肠内分泌功能从而影响全身血糖。

关键词: 肠道菌群, 牙周炎, 糖尿病, 16S rDNA测序

Abstract: Objective: To investigate the impact of Porphyromonas gingivalis (P.g) on blood glucose levels in diabetic mice and identify the bacteria in the intestinal flora closely associated with blood glucose regulation; and to explore the underlying mechanisms of this intervention. Methods: Thirty-two C57BL/6J mice were randomly divided into four groups (n=8 per group) using a randomized control repeated blind method. The groups were as follows: healthy mice fed with phosphate-buffered saline (PBS) (WT group); healthy mice fed with P.g (WT P.g group); diabetic mice injected with streptozotocin and fed with PBS (STZ group); and diabetic mice injected with streptozotocin and fed with P.g (STZ P.g group).The blood glucose levels, body weight, and mental state of the mice were observed weekly. 16S rDNA analysis was performed to assess the composition changes of intestinal flora in colon feces. Immunofluorescence and enzyme-linked immunosorbent assay (ELISA) techniques were used to measure the expression of glucagon-like peptide-1 (GLP-1). Additionally, quantitative polymerase chain reaction (qPCR) was employed to examine the expression of prohormone converting enzyme-1 (PC1/3) and proglucagon mRNA in colon tissue. Results: No significant difference in fasting blood glucose (FBG) levels was observed between the WT group and the WT P.g group. However, at the sixth week after oral administration, a significant difference was observed between the STZ group and the STZ P.g group, with the latter showing higher blood glucose levels. The richness and diversity of the intestinal flora were significantly decreased in the STZ P.g group. qPCR analysis revealed abnormal expression of PC1/3 and proglucagon mRNA, as well as significantly decreased expression of GLP-1. The 16S rDNA analysis indicated a significant increase in the richness of Aeromonas, Rothia, Paraprevotella, and Clostridium in the STZ P.g group, while the richness of Ligilactobacillus, Desulfovibrio, Akkermansia, Bacteroides, and Lachnospiraceae was significantly reduced. FBG levels showed a negative correlation with the richness of Lachnospiraceae_NK4A136_group, Enterorhabdus, and Candidatus_Saccharimonas, but a positive correlation with Desulfovibrio, Akkermansia, and Bacteroides. Conclusion: The results suggest that P.g may interfere with intestinal endocrine function through modulating the intestinal flora, potentially influencing blood glucose regulation throughout the body.

Key words: intestinal microbiota, periodontitis, diabetes, 16S rDNA sequencing

陈文文, 闫福华, 莫朝伦, 黄明坤, 李萧纹, 张军梅. 牙龈卟啉单胞菌通过干扰肠道菌群对糖尿病小鼠血糖的影响[J]. 口腔医学研究, 2023, 39(11): 1005-1011.

CHEN Wenwen, YAN Fuhua, MO Chaolun, HUANG Mingkun, LI Xiaowen, ZHANG Junmei. Effect of Porphyromonas Gingivalis on Blood Glucose and Intestinal Flora in Diabetic Mice: A Mechanistic Exploration[J]. Journal of Oral Science Research, 2023, 39(11): 1005-1011.

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牙龈卟啉单胞菌通过干扰肠道菌群对糖尿病小鼠血糖的影响

Effect of Porphyromonas Gingivalis on Blood Glucose and Intestinal Flora in Diabetic Mice: A Mechanistic Exploration

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